The effect of ligand binding on the proteolytic pattern of methylmalonate semialdehyde dehydrogenase

Natalia Y. Kedishvili, Kirill M. Popov, Robert A. Harris

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Native rat liver methylmalonate semialdehyde dehydrogenase was proteolyzed by lysylendopeptidase C, chymotrypsin, and trypsin to generate different cleavage fragments of molecular masses: 50, 8, 55, 44, 39, 53, 45, and 40 kDa. A proteolytic cleavage map of MMSDH was constructed based on sequencing data and a comparison of appearance and degradation rates of the different protein fragments as shown by SDS-PAGE. NAD+ was highly effective as a protector against proteolysis in both the N-terminal and the C-terminal parts of the intact enzyme. NADH did not efficiently protect the intact enzyme; however, it stabilized proteolytic fragment L50 from further degradation. This suggests that the NAD+-binding domain is not destroyed by cleavage of the N-terminal part of MMSDH. CoA had no effect on the proteolytic cleavage patterns of MMSDH. However, CoA esters reduced the protective effect of NAD+ with an order of effectiveness of acetyl-CoA > propionyl-CoA > butyrylCoA. p-Nitrophenyl acetate, substrate for esterase activity by the enzyme, partially prevented the protective effect of NAD+ against proteolysis. These results suggest that S-acylation of the enzyme prevents a stabilizing conformational change induced in MMSDH by NAD+ binding.

Original languageEnglish (US)
Pages (from-to)21-26
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume290
Issue number1
DOIs
StatePublished - Oct 1991

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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